FIG. 5.

Apoptosis and activation of the caspase pathway by ZD55-TRAIL-IETD-Smac. (A) Bel-7404, NCI-H460, Huh-7, and Becap-37 cells were infected with ZD55-TRAIL-IETD-Smac at an MOI of 10 for 48 hr. Nuclei were stained with Hoechst 33342 (5 μg/ml) to visualize chromatin condensation, nuclear shrinkage, or fragmentation (markers of apoptosis; arrows) by fluorescence microscopy (original magnification,×200). (B) Left: Percent apoptotic cell death was determined by FACS 48 hr after treatment of Bel-7404 and QSG-7701 cell groups with ZD55, ZD55-TRAIL, ZD55-Smac, a combination of ZD55-TRAIL and ZD55-Smac (at a ratio of 1:1), and ZD55-TRAIL-IETD-Smac at 10 MOI. Right: A column drawing representing the mean values of three cytometric assays. **p<0.01 versus ZD55-TRAIL and ZD55-Smac. (C) Bel-7404 cells were infected with ZD55-TRAIL-IETD-Smac and the other adenoviruses for 48 hr at the dose described previously. Whole-cell extracts were prepared and immunoblotted to detect activation of the caspase pathway. β-Actin was used as a loading control. The data are representative of two determinations with identical results. PARP, poly(ADP-ribose) polymerase. Color images available online at www.liebertpub.com/hum